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O. Grüter, C. Kostic, M. Tekaya, D.F. Schorderet, L. Zografos, F.L. Munier, Y. Arsenijevic; Potential Improvement of Lentiviral Gene Transfer by Weakening The Extracellular Matrix . Invest. Ophthalmol. Vis. Sci. 2004;45(13):4773.
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Purpose: Gene transfer seems to offer a substantial promise for the therapy of degenerative ocular diseases. Lentiviral vectors have the ability to transduce efficiently murine photoreceptors (PR) during the first days of life, but they are poorly effective on PR during adulthood (Kostic et al. (2003)). Previous studies have described the abilities of enzymes (chondroitinase ABC (CA) and neuraminidase (NM)) to change the structure of the interphotoreceptor matrix (IPM) and to increase the retinal detachment when subretinally injected without affecting retinal function after recovery (Yao et al. (1992)) . Considering the IPM as a physical barrier that may decrease photoreceptor transduction, we injected into the subretinal space of the mouse different enzymes simultaneously with the lentiviral vector preparation to increase the viral transduction by fragilizing the IPM. Methods: NM or CA and the lentiviral vector LV–Rho–GFP expressing GFP under the control of the rhodopsin promoter were simultaneously injected into the subretinal space of adult DBA mice. The animals were sacrificed 7 days post–injection and eyes were processed by cryostat sectioning and analysed by fluorescent microscopy. Results: Subretinal injection of NM and CA induces modifications in the IPM of the mouse as previously described in the rabbit (Yao et al. (1990)). The simultaneous subretinal injection of NM and LV–Rho–GFP increases markedly and significantly the number of transduced PR (more than 4 fold) and is correlated with an increase of the retinal detachment. CA effect has a similar tendancy but the increase is not statistically significant. Conclusions: The lentiviral transduction is improved when simultaneously injected into the subretinal space of the mouse with the NM. Among the enzyme properties, the increase of the retinal detachment appears to facilitate the lentiviral transduction. This technique seems to be very promising for increasing the lentiviral vector diffusion into the subretinal space. Nevertheless, the transduction density is not increased at a precise point of the retina and complementary studies of the IPM should be made to identify and better understand the factor which limits the PR transduction by lentiviral vectors.
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